caa a22 4500 397494300 CHVBK 20180308164517.0 cr unu---uuuuu 161202e199503 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a124762 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a124762 Photoaffinity Labeling of Influenza Virus RNA Polymerase PB1 Subunit with 8-Azido GTP [Elektronische Daten] [Yukiyasu Asano, Kiyohisa Mizumoto, Tokumi Maruyama, Akira Ishihama] 8-Azido GTP (8-N3 GTP) was demonstrated to be polymerized into RNA by influenza virus-associated RNA polymerase at about one tenth the rate of GTP incorporation. The Km value for the azido analogue of GTP in primer-dependent RNA synthesis was 94 μM whereas Km for the natural substrate, GTP, was 6.7 μM. Upon exposure of a mixture of 8-N3 [α-32P]GTP and influenza virus ribonucleoprotein (RNP) complexes to ultraviolet light, the PB1 subunit of viral RNA polymerase was selectively radio-labeled. The photo-labeling of PB1 was competed strongly by GTP and to lesser extents by other nucleoside 5′-triphos-phates. These results altogether support the prediction that the substrate-binding site (S site) of influenza RNA polymerase is located on the PB1 protein. In the presence of ApG primer, the 8-N3 GTP binding was reduced to about 40% level, suggesting that the GTP analogue can bind not only to the S site but also to the primer- and product-binding site (P site). © 1995 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence 8-azido guanine nucleotide nationallicence photoaffinity cross-linking nationallicence RNA synthesis nationallicence subunit function nationallicence transcriptase-replicase nationallicence Asano Yukiyasu Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut Mizumoto Kiyohisa Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut Maruyama Tokumi Faculty of Pharmaceutical Sciences, Tokushima Bunri University Tokushima, Tokushima 770 aut Ishihama Akira Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut The Journal of Biochemistry Oxford University Press 117/3(1995-03), 677-682 0021-924X 117:3<677 1995 117 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a124762 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a124762 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Asano Yukiyasu Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut NATIONALLICENCE 700 1- Mizumoto Kiyohisa Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut NATIONALLICENCE 700 1- Maruyama Tokumi Faculty of Pharmaceutical Sciences, Tokushima Bunri University Tokushima, Tokushima 770 aut NATIONALLICENCE 700 1- Ishihama Akira Department of Molecular Genetics, National Institute of Genetics Mishima, Shizuoka 411 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 117/3(1995-03), 677-682 0021-924X 117:3<677 1995 117 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford